Control of parasitic mites of honey bees

ABSTRACT

The present invention is directed to methods and compositions for use to control parasitic mites of honey bees, particularly Varroa mites. In one aspect, the invention is directed to control of parasitic mites of honey bees wherein the active ingredient is a miticidally effective amount of a selected ketone or 1-heptanol, ethyl butyrate, benzaldehyde, heptaldehyde, or d-limonene. In a second aspect, the invention is directed to control of parasitic mites of honey bees wherein the active ingredient is an effective attractant amount of 2-heptanone. The attracted mites are then trapped or otherwise removed from the locus of the bees. The present invention is also directed to methods and compositions which include 2-heptanone to control hive invading pests of honey bees.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. ProvisionalApplication No. 60/272,097 filed Feb. 28, 2001. The disclosure of saidprovisional application is incorporated herein by reference in itsentirety.

BACKGROUND OF THE INVENTION

[0002] 1. Field of the Invention

[0003] The present invention is directed to methods and compositions tocontrol parasitic mites of honey bees and methods and compositions tocontrol hive invading pests.

[0004] 2. Description of the Art

[0005] Honey bees are necessary to pollinate important agriculturalcrops and also to produce honey and wax for commercial markets. In theUnited States, honey bees produce $270 million worth of honey, beeswax,and other hive products and pollinate over $14 billion worth of cropsannually.

[0006] Parasitic mites are economically important parasites of honeybees which affect honey bee populations worldwide. Varroa jacobsoni(Oudemans) (=V. destructor) (Acari: Varroidae) are small crab-shapedparasitic mites that are found attached to adult bees or under cappedbrood cells where they reproduce. In either case, Varroa feed onhemolymph by puncturing the exoskeleton of the bee with theirmouthparts.

[0007] Varroa reproduction begins when the adult female enters a broodcell shortly before it is capped. The female must feed on larvalhemolymph before she can lay eggs. The Varroa eggs eclose under thesealed cell, and the developing mites feed on the bee pupa. The firstegg laid by the female Varroa develops into a male. Subsequent eggsdevelop into females that mate with their brother. The mated femalemites along with their mother are released from the capped cell when thebee emerges. These mites will hereinafter be referred to as “phoreticmites.” The females attach to adult bees between the abdominal segmentsor between body regions, making them difficult to detect. These are alsoplaces from which they can easily feed on the bees' hemolymph. Adultbees serve as intermediate hosts when little or no brood is availableand as a means of transport.

[0008] Varroa mites reduce bee longevity. When infestation levels arehigh, entire colonies die. Varroa are also believed to damage honey beesdue to transmission of at least six bee viruses. Spread of Varroa mitesamong colonies can occur due to several factors, including commercialtransport of bees and queens, the migratory activities of beekeepers orswarms that may fly long distances.

[0009] Maintaining a supply of strong honey bee colonies available forpollination is essential for the sustained production of crops worthmore than $14 billion to the U.S. farm economy. Current control measuresfor Varroa include synthetic pesticides. Introducing lipophilicpesticides into honey bee colonies not only contaminates the comb, butopens the possibility of contamination of honey and pollen which aresold to the public.

[0010] Presently there is one EPA-registered product under Section 3 ofthe Federal Insecticide, Fungicide and Rodenticide Act (FIFRA) forcontrol of Varroa mites: a plastic strip impregnated with the contactsynthetic pyrethroid pesticide fluvalinate (Apistan® strip, WellmarkInternational). There is also one EPA registered product under Section 3of FIFRA for suppression of Varroa mites: a 65% formic acid gel(Apicure®, Apicure, Inc., believed to be the device of U.S. Pat. No.6,037,374). In 1999 and 2000 EPA issued an emergency authorization,under Section 18 of FIFRA, for use of the organophosphate coumaphos(CheckMite+™, Bayer Corp.) for control of Varroa mites. Formic acid andcoumaphos are in EPA toxicity category I which requires the signal word“Danger.” Fluvalinate is in EPA toxicity category III which requires asignal word of “Caution.” 2-Heptanone is also in toxicity category III.

[0011] Of serious concern is the fact that Varroa mite resistance tofluvalinate, the most commonly used miticide, has been reported inEurope (Milani, Apidologie 30:229-234 (1999); Vedova et al., Ape NostraAmica 19:6-10 (1997)) and in the United States (Elzen et al., AmericanBee Journal 138:674-676 (1998); Elzen et al., Apidologie 30:13-18(1999)). Also, fluvalinate residues have been detected in foundationbeeswax used in bee colonies. Resistance to coumaphos is also now beingreported. Further, coumaphos, which is also lipophilic, poses the threatof contaminating hive product. The formic acid compositions areeffective against tracheal mites but are reported to have limitedeffectiveness against Varroa. Packaging problems have been reported forthe formic acid compositions.

[0012] To avoid some of the potential problems related to pesticide usein beekeeping, some researchers are selecting strains of bees thattolerate or are resistant to mites. These selection programs takeseveral years, and those colonies that are susceptible to mites will belost in the interim. Beekeepers need immediate relief from Varroainfestations that already exist in their colonies. What is needed aresafe and effective ways to control parasitic mites of bees.

[0013] Hive invading pests inflict substantial economic losses tobeekeepers. One of the most damaging pests to the bee industry is thegreater wax moth (Galleria mellonella). Currently there is no registeredmeans of controlling this pest. The small hive beetle (Aethina tumida),is another destructive pest. There are no known natural enemies of thesmall hive beetle in the United States. As discussed below, other hiveinvading pests include ants and the parasitic mite Tropilaelaps. What isneeded are safe and effective ways to control hive invading pests.

[0014] 2-Heptanone is a pheromone produced by the mandibular glands ofadult worker honey bees, Apis mellifera and Apis cerana (Vallet et al.,J. Insect Physiol. 37(11):789-804 (1991); Sakamoto et al., Journal ofApiculture Research 29(4):199-205 (1990)), older than 8-10 days. Theopening of the mandibular gland is inside the buccal cavity (mouth) ofthe bee at the base of the mandibles. 2-Heptanone is producedcontinuously and is universally distributed throughout the bee colonyand in the wax. It is believed that the primary function of 2-heptanonein the honey bee hive is that of the principal universal solvent used bythe bees to manufacture bees wax comb and propolis (bee glue used tosuspend wax combs and plug holes). The bees secrete 2-heptanone whilethey use their mandibles to masticate (chew) the tiny wax flakesproduced by their abdominal wax glands. The wax flakes are formed intouniformly thin wax sheets that are used to build the solid hexagonal waxwalls of honey comb cells. Worker bees also gather a variety of plantresins which are solubilized with 2-heptanone and either painted on thesurface of the wax honey comb or mixed with wax to produce propolis. Anew layer of propolis is painted in brood cells during cleaning aftereach brood cycle. The net result is that 2-heptanone is incorporatedinto the structure of the hive interior.

SUMMARY OF THE INVENTION

[0015] The present invention is directed to methods and compositions tocontrol parasitic mites of honey bees and methods and compositions tocontrol hive invading pests.

[0016] In one embodiment, the invention is directed to miticidalcompositions and use thereof to control parasitic mites of honey bees,wherein the active ingredient of the composition is a miticidallyeffective amount of a compound, which comprises:

[0017] (a) a ketone of the structure:

CH₃—(CH2)_(x)—CO—(CH2)_(y)—CH₃

[0018]  wherein y is 0 and x is 0 to 5 or wherein y is 1 or 2 and x is 3or 2, respectively; or

[0019] (b) a compound selected from the group consisting of 1-heptanol,ethyl butyrate, benzaldehyde, heptaldehyde, and d-limonene.

[0020] In a second embodiment, the invention is directed to attractantcompositions and use thereof to control parasitic mites of honey bees,wherein the active ingredient of the composition is an effectiveattractant amount of 2-heptanone. In this embodiment, mites are exposedto the 2-heptanone, and the attracted mites are then trapped orotherwise removed from the locus of the bees. The composition can beused to detect, survey, monitor, or diagnose mite levels, and controlmites by trapping out or otherwise incapacitating attracted mites.

[0021] The invention is also directed to methods and compositionscomprising 2-heptanone to control hive invading pests. In thisembodiment, a selected area, such as a bee hive, bee equipment and thelike, is exposed to an effective hive invader-controlling amount of2-heptanone to prevent hive invading pests from invading the area or toreduce the numbers of pests invading the area.

[0022] Currently, there is no economically feasible means to determineif a colony has Varroa mites. Since Varroa can enter a colony onincoming bees, the mite can be present in a colony at any time.Determining if a colony has Varroa is essential for evaluating ifcontrol measures are required. Because Varroa can enter colonies at anytime when bees are foraging, there is not a particular time of year whenVarroa might be found in colonies. Once Varroa establish in a colony,they must be treated to avoid devastating effects on colony populationsparticularly at certain times of year. The attractant embodiment of theinvention provides a simple inexpensive means to monitor the presence ofVarroa in colonies and provides a pro-active way to reduce the number ofmites infesting mite-free colonies.

[0023] In accordance with this discovery, it is an object of theinvention to provide methods and compositions for control of parasiticmites of honey bees, particularly Varroa mites, and to provide methodsand compositions for control of hive invading pests.

[0024] Another object of the invention is the provision of safe andeffective mite control methods and compositions as alternatives tosynthetic pesticides (e.g., fluvalinate and coumaphos) or formic acid.

[0025] With regard to 2-heptanone, since this compound already exists inbee hives at low levels, there is little chance of toxicity to bees orcontamination of hive products.

[0026] The miticidal compounds of the invention can be used as aprophylactic to maintain parasitic mite infestation levels below theeconomic threshold, or reactively to control infestation outbreaks ofmites in honey bee colonies. The miticidal compounds can be used aloneor in conjunction with other measures incorporated into an IntegratedPest Management Program.

[0027] A further object of the invention is to provide miticides fordirect control of parasitic mites of honey bees. In one aspect, theinvention provides control using 2-heptanone, a compound that naturallyoccurs in the colony, and thus, the invention is useful to insure thehealth and vigor of honey bee colonies available for pollination andinsure an affordable food supply. In one aspect of the invention,naturally occurring levels of 2-heptanone are augmented to ensurepersistence of this volatile compound at miticidal levels.

[0028] A still further object of the invention is to provide a means forattracting and trapping Varroa mites that are searching for honey beelarvae or to attract and trap mites that have attached to robber bees toprevent the mites from infesting new colonies.

[0029] An even further object of the invention is to provide a trappingdevice using 2-heptanone in an attractant amount to provide inexpensiveand effective means to monitor Varroa mites while not disrupting honeybee colony behavior or vigor.

[0030] It is also an object of the invention to provide a means forcontrolling hive invading pests.

[0031] Other objects and advantages of the invention will become readilyapparent from the ensuing description.

DETAILED DESCRIPTION OF THE INVENTION

[0032] The present invention is directed to methods and compositions tocontrol parasitic mites of honey bees and methods and compositions tocontrol hive invading pests. In one embodiment, the invention isdirected to methods and compositions wherein selected compounds areeffective miticides for control of parasitic mites of honey bees. In asecond embodiment, the invention is directed to methods and compositionscomprising 2-heptanone as an attractant for controlling mites. Inanother embodiment, 2-heptanone is used for control of hive invadingpests.

[0033] Definitions:

[0034] Unless defined otherwise, all technical and scientific terms usedherein have the meaning commonly understood by a person skilled in theart to which this invention belongs. The following reference providesone of skill with a general definition of many of the terms used in thisinvention: The Hive and The Honeybee, Ed. Joe M. Graham, Dadant & Sons,Inc., Hamilton, Ill. 1993.

[0035] To facilitate understanding of the invention, a number of termsare defined below.

[0036] 2-Heptanone, CH₃(CH₂)₄COCH₃, CAS Registry No. 110-43-0, alsoknown as methyl amyl ketone and methyl pentyl ketone, is a volatileliquid at room temperature [d₄ ¹⁵ 0.8197; b.p.₇₆₀ 151.5° C.]. It issoluble in alcohol or ether and very slightly soluble in water. It isavailable commercially.

[0037] Acetone, CH₃COCH₃, CAS Registry No. 67-64-1, also known as2-propanone and dimethyl ketone, is a volatile liquid at roomtemperature [d₂₅ ²⁵ 0.788; b.p. 56.5° C.]. It is miscible with water,alcohol, dimethylformamide, chloroform, ether, and most oils. It isavailable commercially.

[0038] 2-Butanone, CH₃CH₂COCH₃, CAS Registry No. 78-93-3, also known asmethyl ethyl ketone, is a volatile liquid at room temperature [d₄ ²⁰0.805; b.p. 79.6° C.]. It is soluble in about four parts water andmiscible with alcohol, ether, and benzene. It is available commercially.

[0039] 2-Pentanone, CH₃(CH₂)₂COCH₃, CAS Registry No. 107-87-9, alsoknown as methyl propyl ketone, is a liquid at room temperature [d₄ ²⁰0.809; b.p. 102° C.]. It is almost insoluble in water and is misciblewith alcohol or ether. It is available commercially.

[0040] 2-Hexanone, CH₃(CH₂)₃COCH₃, CAS Registry No. 591-78-6 is a liquidat room temperature [b.p. 127° C.]. It is available commercially.

[0041] 2-Octanone, CH₃(CH₂)₅COCH₃, CAS Registry No. 111-13-7, also knownas methyl hexyl ketone, is a liquid at room temperature [b.p.173° C.].It is available commercially.

[0042] 3-Heptanone, CH₃(CH₂)₃COCH₂CH₃, CAS Registry No. 106-35-4, alsoknown as butyl ethyl ketone, is a liquid at room temperature [b.p.146-149° C.]. It is available commercially.

[0043] 4-Heptanone, CH₃(CH₂)₂CO(CH₂)₂CH₃, CAS Registry No. 123-19-3,also known as dipropyl ketone, is a liquid at room temperature [b.p.145° C.]. It is available commercially.

[0044] 1-Heptanol, CH₃(CH₂)₅CH₂OH, CAS Registry No. 111-70-6, also knownas n-heptyl alcohol, is a volatile liquid at room temperature [d₄ ²⁵0.8187; b.p.₇₆₀ 175.8° C.]. It is miscible with alcohol or ether. It isavailable commercially.

[0045] Ethyl butyrate, CH₃CH₂CH₂COCH₂CH₃, CAS Registry No. 105-54-4,also known as butanoic acid ethyl ester, butyric acid ethyl ester, andethyl n-butyrate, is a liquid at room temperature [d₄ ²⁰ 0.879; b.p.120-121° C.]. It is soluble in about 150 parts water; miscible withalcohol or ether. It is available commercially.

[0046] Benzaldehyde, C₇H₆O, CAS Registry No. 100-52-7, also known asbenzoic aldehyde and artificial essential oil of almond, is a liquid atroom temperature [d₄ ¹⁵ 1.050; b.p. 179° C.]. It is soluble in about 350parts water; miscible with alcohol or ether. It is availablecommercially.

[0047] Heptaldehyde, CH₃(CH₂)₅CHO, CAS Registry No. 111-71-7, also knownas heptanal, is a liquid at room temperature [b.p. 153° C.]. It isavailable commercially.

[0048] d-Limonene, C₁₀H₁₆, CAS Registry No. 5989-27-5, also known as(R)-(+)-limonene, (+)-4-isopropenyl-1-methyl-1-cyclohexene and(+)-p-mentha-1,8-diene, is a liquid at room temperature [d₄ ²¹ 0.8402;b.p.₇₆₃ 175.5-176° C.]. It is available commercially.

[0049] The term “honey bee” refers to members of the Order Hymeoptera,Family Apidae and includes by way of example, the species Apismellifera, and Apis cerana.

[0050] The term “colony” refers to a community of bees with a singlequeen, thousands of workers, and brood. During part of the year thereare also several hundred drones. The bees live and work together as onefamily in a hive.

[0051] The term “comb” refers to sections of hexagonal bees wax cellsbuilt by honey bees and used to rear their brood and store honey andpollen. The cells in each comb are built back-to-back with a commoninterior wall. The combs are arranged in parallel series.

[0052] The term “hive” refers to the cavity/domicile occupied by a honeybee colony. The modern box hive includes a bottom board, cover, and oneor more boxes, stacked one above the other. Inside, each box contains aseries of movable frames of comb or foundation held in a verticalposition a bee space apart.

[0053] For the purposes of this invention, a compound is applied for itsintended purpose at a level that is greater than the ambient backgroundlevel. This is described further with reference to 2-heptanone and itsuse as a miticide, but applies to all embodiments of the invention. Asdiscussed above, 2-heptanone is produced by honey bees, and thus isnaturally present in the ambient air of a colony, denoted hereinafter asthe “background level.” For the purposes of this invention, theeffective amount of 2-heptanone to control mites is an amount greaterthan the ambient background level of 2-heptanone naturally present inthe air at the time of exposure. That is, control of parasitic mites iscarried out by exposing the target mites to a source of 2-heptanoneother than or in addition to the naturally present background level. Inone aspect of the invention, naturally occurring levels of 2-heptanoneare augmented to ensure persistence of this volatile compound atmiticidal levels or at hive invader-controlling levels.

[0054] The background amount of 2-heptanone in a hive depends on factorssuch as time of year, colony size, amount of comb being built, amount ofbrood in the colony, temperature, and colony demographics (agedistribution in the colony). Vallet et al., 1991, supra, estimated theamount of 2-heptanone in a honey bee mandibular gland to be from about0.1 microliter at emergence to 7 microliters in foraging bees.Determination of the ambient background level of 2-heptanone in the airin a hive (or other locus) of honey bees in a particular set ofcircumstances can be determined by sampling the air and analyzing forthe amount of 2-heptanone per volume of air space using standard GC/MSanalysis.

[0055] Control of parasitic mites of honey bees refers to any method ormeans using 2-heptanone or other compound specified above thateliminates or reduces the numbers of mites available to affect honeybees. This includes removing mites from an area where bees are presentor may occur; preventing mites from advancing to mite-free areas orreducing the number of mites from advancing to mite-free areas. Controlof parasitic mites also includes any method or means using 2-heptanoneto attract mites so that they can be trapped or otherwise removed fromthe locus of the bees.

[0056] Control of hive invading pests refers to any method or meansusing 2-heptanone that eliminates or reduces the numbers of hiveinvading pests available to affect honey bees.

[0057] Dispensing means refers to any means for dispensing controllingamounts of 2-heptanone or other compound specified above. For purposesof this invention, a dispensing means, in its broadest ambit, is definedas any means which both (a) contains or holds unvolatilized compound and(b) releases the compound into the air.

[0058] A dispensing means may take several forms. In general, adispensing means will comprise a means for holding an amount of thecompound within a space and for release into the atmosphere. Suchdispensing means may be solid or liquid devices or formulations such asmonolithic systems, laminated structures, and reservoir systems with orwithout rate-controlling membranes or formulations. For example, adispensing means may be as simple as a reservoir or an adsorbent orabsorbent material such as cotton or paper, which dispensing means bothholds and releases the compound. A preferred dispensing means is adevice or formulation which provides controlled release, slow release orsustained release of the compound, as discussed in detail below.

[0059] Several types of controlled-release devices exist: those in whichthe active ingredient forms a core surrounded by an inert diffusionbarrier (such devices are frequently referred to as “reservoir”devices); and those in which the active ingredient is dissolved ordispersed in an inert diffusion barrier (such devices are frequentlyreferred to as “monolithic” devices). Each of these devices canobviously exist in a variety of shapes, and can be degradable ornon-degradable. Sustained release can also be achieved by a number ofother methods such as complexation of the active ingredient, slowlydissolving coatings, erosion, microbial action, use of derivatives ornew compounds of reduced solubility or volatility, and the likes.

[0060] In monolithic devices, the active ingredient is dispersedthroughout (or dissolved in) a substantially inert matrix from which theactive ingredient is gradually released in the environment. Non-limitingexamples of matrices that have been included in monolithic devicesinclude various gels, waxes, gelatins, natural resins, rubbers,elastomers, synthetic and natural polymers, and the likes. In reservoirdevices, several classes of devices exist. One important class includesmembranes which are non-porous, homogeneous polymeric films, throughwhich transport occurs by a process of dissolution of the permeatingspecies in the polymer at one interface and diffusion down a gradient inthermodynamic activity. These membranes are usually referred to assolution-diffusion membranes. Another class includes the porous and/orfibrous barriers such as, for example, hollow fibers, porous and/orfibrous materials, in which the active ingredient diffuses mainly bycapillary forces. Other less common devices exist where diffusion istaking place under external forces (e.g., gravity, electrical field,vacuum, centrifugal forces, etc.) or mechanical pumping, and the likes.

[0061] Formulations such as microencapsulations and emulsions can beused to slow down the release of the active ingredient. Emulsionformulations can be found as water in oil (w/o) or oil in water (o/w).Droplet size can vary from the nanometer scale (colloidal dispersion) toseveral hundred microns. A variety of surfactants and thickeners areusually incorporated in the formulation to modify the size of thedroplets, stabilize the emulsion, and modify the release. Microcapsulesare small particles that contain a core material or active ingredientsurrounded by a coating or shell. Size typically varies from 1 to 1000microns with capsules smaller than 1 micron classified as nanocapsulesand capsules larger than 1000 microns as macrocapsules. Core payloadusually varies from 0.1 to 98 weight percent. Microcapsules can have avariety of structures (continuous core/shell, multinuclear, ormonolithic) and have irregular or geometric shapes. Several processesfor preparing microcapsules are described in the literature.Encapsulation processes are often loosely classified as either chemicalor mechanical. Examples of chemical processes include but are notlimited to complex coacervation, polymer-polymer incompatibility,interfacial polymerization in liquid media, in situ polymerization,in-liquid drying, thermal and ionic gelation in liquid media,desolvation in liquid media, starch-based chemistry processes, trappingin cyclodextrins, and formation of liposomes. Examples of mechanicalprocesses include but are not limited to spray drying, spray chilling,fluidized bed, electrostatic deposition, centrifugal extrusion, spinningdisk or rotational suspension separation, annular-jet encapsulation,polymerization at liquid-gas or solid-gas interface, solventevaporation, pressure extrusion or spraying into solvent extractionbath.

[0062] Without being limiting, specific exemplary solid controlledrelease substrates include porous particulates or substrates such assilica, perlite, talc, clay, pyrophyllite, diatomaceous earth, gelatinand gels, polymers (e.g., polyurea, polyurethane, polyamide, polyester,etc.), polymeric particles, or cellulose. These include, for example,hollow fibers, hollow tubes or tubing which release 2-heptanone or othercompound specified above through the walls, capillary tubing whichreleases the compound out of an opening in the tubing, polymeric blocksof different shapes, e.g., strips, blocks, tablets, discs, which releasethe compound out of the polymer matrix, membrane systems which hold thecompound within an impermeable container and release it through ameasured permeable membrane, and combinations of the foregoing. Examplesof other dispensing means are polymer laminates, polyvinyl chloridepellets, and microcapillaries. Another dispensing means includes usingmicroencapsulation techniques to encapsulate the compound. Thisincludes, for example, encapsulation of the compound in a polyvinylchloride (PVC)-polyvinyl acetate (PVA) plastic (see for example, Riethet al., Journal of Apiculture Research 25(2):78-84 (1986)). A dispensermay also comprise a release substrate, with the release into theatmosphere controlled by a permeable wall or membrane or by a smallopening surrounded by an impermeable wall or membrane or a chemicalcomposition such as a gel composition which holds and releases thecompound. Examples of acrylic block controlled release formulations,controlled release strip formulations, and microencapsulationformulations are described in further detail, below and in the Examples.

[0063] Liquid forms of release substrates include vegetable and/ormineral oils, preferably containing surface active agents to render thecomposition readily dispersable in water, such agents include wettingagents, emulsifying agents, dispersing agents, and the like.

[0064] Dispensing means for controlled release are described in U.S.Pat. Nos. 5,750,129; 4,775,534; 5,849,317; 6,037,374; 3,577,515, whichare incorporated herein by reference in their entirety.

[0065] Miticides of the Invention

[0066] In this embodiment, the invention is directed to miticidalcompositions and use thereof to control parasitic mites of honey bees.The miticidal compounds of the invention comprise:

[0067] (a) ketones of the structure:

CH₃—(CH2)_(x)—CO—(CH2)_(y)—CH3

[0068]  wherein y is 0 and x is 0 to 5 or wherein y is 1 or 2 and x is 3or 2, respectively; or

[0069] (b) compounds selected from the group consisting of 1-heptanol,ethyl butyrate, benzaldehyde, heptaldehyde, and d-limonene.

[0070] The miticidal compositions contain as the active ingredient oneor more of the miticidal compounds of the invention in an effectivemiticidal amount. This embodiment is useful to control free running orphoretic mites in bee colonies or other areas in which an effectiveamount can be maintained.

[0071] In this aspect, the term control includes treating mites with aneffective amount of miticidal compound, that is, an amount which iseffective to kill mites, to incapacitate mites such as by disruptingneural or other physiological functions to prevent essential mitefunctions or reproduction, or render mites impaired sufficiently to betrapped, drowned, isolated, or otherwise removed from an area. Aneffective miticidal amount is determined as that quantity of compoundemitted from a formulation or dispenser holding the compound that issufficient to accomplish the aforenamed control activities. Treatmentmay also comprise augmenting naturally occurring levels of 2-heptanoneto ensure persistence of this volatile compound at effective levels.

[0072] The quantity of miticidal compound must also be insufficient topermanently remove bees from a colony. Tests to determine this can bereadily carried out as described in Example 5, below. In our tests ofthe effects of 2-heptanone on honey bee colony behavior (see Example 5)we found that even very high concentrations tested of 2-heptanone in acolony did not, at any time, cause all bees in the colony to fully andpermanently exit the hive. Additionally, our data from colonies andobservation hives indicate that effective miticidal concentrations of2-heptanone do not affect the worker bees in relation to their behaviortoward the queen, queen egg laying, or the tendency of bees to remain inthe hive and care for the brood.

[0073] For control of parasitic mites, particularly Varroa mites, in beecolonies, it is preferred that treatment be carried out over a completebrood cycle (21 days for workers or 28 days for drones) and morepreferably two or more brood cycles, that is, 42 days or longer.

[0074] It is envisioned that the composition of the invention would beuseful in killing or incapacitating mites when used with any dispensingmeans, as described in detail, above. Conveniently, a dispensing meansmay be left in the hive until all the miticidal compound evaporates.

[0075] Examples of dispensing means include a reservoir, controlledrelease acrylic block, or controlled release strip as described inExamples, below. In any particular instance, the optimum formulationwill depend on the mold shape and the active ingredient proportion.

[0076] An exemplary acrylic block controlled release formulation isprepared as follows: 5% to 80% of 2-heptanone is mixed with 20% to 95%isobornyl methacrylate and 0% to 30% tripropyleneglycol diacrylate.After the mixing step, benzoyl peroxide is introduced within the mixerat a ratio between 0.05% and 5% and left for homogenization during 30minutes. Then dimethylparatoluidine (0.05% to 5%) is introduced into themixer, and the polymerization starts at room temperature. The mixture isintroduced into a mold. Polymerization proceeds during several hours andwhen the temperature of the device is cooled down, the device is removedfrom the mold.

[0077] A more preferred acrylic block formulation comprises: 30% to 70%of 2-heptanone; 30% to 70% of isobornyl methacrylate; 1% to 20%tripropylene diacrylate; 1% to 4% benzoyl peroxide; 0.5% to 2%dimethylparatoluidine. The most preferred formulation comprises: 50% to60% of 2-heptanone; 30% to 40% of isobornyl methacrylate; 5% to 10%tripropylene diacrylate; 1% to 3% benzoyl peroxide; 0.5% to 2%dimethylparatoluidine.

[0078] A controlled release strip formulation can be composed of a solidelastomer matrix impregnated by the compound, e.g., 2-heptanone, andsurrounded by a permeable release rate controlling membrane, such asdescribed in U.S. Pat. No. 5,750,129. An exemplary slow release deviceis as follows: the strip is composed of a polymer sleeve 20 cm long and5 cm wide in which a polyurethane core containing 60 g 2-heptanone wassealed. The controlled diffusion of 2-heptanone through the membrane canoccur over a period of 3 weeks or greater depending on the formulation.

[0079] An exemplary slow release oil-gelled 2-heptanone formulation isas follows: 2-heptanone 10.0% Gelled mineral oil (Versagel ™ C HP) 90.0%

[0080] Formulation is carried out by quickly mixing both compounds underheat before cooling down the mixture to get a clear gelled mass with astrong 2-heptanone odor. Viscosity as well as release of the finalformulation can be modified by using hydrocarbon gels with differentproperties (e.g., Versagel™ F or M series from Penreco) and differentconcentrations in 2-heptanone. Versagel™ is a product of Penreco, aPennzoil/Conoco Partnership, Karns City, Pa. Gel formulations can bedispensed in syringes or guns.

[0081] An exemplary slow release gelatin-based 2-heptanone formulationis as follows: 2-heptanone 23.26% Gelatin (Dynagel) 3.07%Polyvinylalcohol 0.29% Water 73.38%

[0082] An emulsion of 2-heptanone in water was made usingpolyvinylalcohol as surfactant. The emulsion was then mixed with thegelatin, heated and stirred up to complete dissolution. Upon cooling,the molded formulation gave a compact rubber-like device with slowrelease properties.

[0083] Other compounds and materials may be added to a formulationprovided they do not substantially interfere with the miticidal activityof the miticide of the invention. Whether or not an additivesubstantially interferes with the miticidal activity can be determinedby standard test formats, involving direct comparisons of efficacy ofthe miticidal compound without an additive and the miticidal compoundwith an additive. Reductions in miticidal activity may be determinedwith standard statistical analyses.

[0084] The dispensing devices can be located as deemed appropriate bythe beekeeper for any particular set of circumstances, including nextto, within or in contact with the brood nest. For example, without beinglimiting, one or more dispensing means is placed either on the top boardof honey bee colonies or between brood frames. Alternatively, thedispensing means can be placed on the bottom board. Combinations of theforegoing are also contemplated by this invention.

[0085] The miticidal compounds of the invention can be used as aprophylactic to maintain parasitic mite infestation levels below theeconomic threshold, or reactively to control infestation outbreaks ofmites in honey bee colonies. The miticidal compounds can be used aloneor in conjunction with other measures incorporated into an IntegratedPest Management Program. The compounds may also be useful in controllingcertain infectious diseases of honey bees which are carried by mites.

[0086] Use of 2-Heptanone as an Attractant to Control Parasitic Mites ofHoney Bees

[0087] In a second embodiment, control of parasitic mites is carried outusing 2-heptanone as an attractant. An effective attractant amount of2-heptanone is provided in an area to which mites are to be attracted.This includes for example, the brood area which is where mites emergefrom cells and search for new hosts, or selected areas where detecting,surveying, monitoring, diagnosing, and/or controlling of mites isdesired. Attracted mites respond to 2-heptanone present in air, and theymove toward the source of the 2-heptanone. Conveniently, 2-heptanone isdispensed within or adjacent to a trapping means to attract and trap themites.

[0088] Without being bound by theory, it is believed that this mode ofaction relates to putative host seeking behavior of Varroa mites basedon the chemistry (2-heptanone content) of honey bee comb containingbrood of the appropriate age for parasitism. This discovery has led tofeasibility of using 2-heptanone to attract Varroa mites in honey beecolonies for the purposes of diagnosis, surveying, monitoring, andcontrol.

[0089] As envisioned, a sticky or pitfall trap system utilizing2-heptanone as the attractant could be used both to diagnose Varroainfestation levels, and to reactively trap out mites in honey beecolonies wherein control procedures are warranted. An advantage of thedevelopment of a 2-heptanone-based mite trapping system would bereduction of the use of pesticides to diagnose or control Varroa. Anadvantage of the 2-heptanone trapping system is its use as a diagnosticaid, helpful in determining the magnitude of a Varroa infestation andthus the need for treatment. A 2-heptanone trapping system may also beuseful in diagnosing and controlling other in hive parasites such as thesmall hive beetle.

[0090] The 2-heptanone attractant is also useful pro-actively to preventor reduce the numbers of Varroa infesting previously Varroa-freecolonies. Colonies become infested when worker bees fly out and robweakened infested hives. In the infested hive, the mites attachthemselves to the robber bees and ‘hitchhike’ back to the robber beeparent colony where they dismount when near bee brood. A 2-heptanoneattractant dispenser would be useful to cause the mites to dismount atsuch a distance from the brood area of the parent colony that the miteswould perish before they could encounter and parasitize bee brood.

[0091] An effective attractant amount of 2-heptanone is provided in anarea to which mites are to be attracted. An effective attractant amountis defined as that quantity of 2-heptanone that attracts mites to thelocation of the 2-heptanone at a rate higher than mites are attracted toa location devoid of the added 2-heptanone attractant. An effectiveattractant amount is determined as the quantity of 2-heptanone emittedfrom a formulation or dispenser holding 2-heptanone that is sufficientto elicit an attraction response from mites.

[0092] It is envisioned that the attractant of the invention would beuseful in detecting, surveying, monitoring, diagnosing or controllingmites when used as a lure. A lure includes a dispenser means whichcontains the attractant. As discussed above, a dispenser means isdefined as any means which both (a) contains or holds unvolatilized2-heptanone and (b) releases 2-heptanone into the air. A dispensingmeans may take several forms as discussed in detail above, andincorporated herein by reference.

[0093] In use in bee colonies, the release device or formulationcontaining an attractant amount of 2-heptanone can be convenientlyplaced between brood frames in honey bee colonies. The device attractsmites such as Varroa that are searching for honey bee larvae in cellsthat are just about to be sealed. Traps are used by inserting them intocolonies. After a selected period of time, e.g., 48 hours after placingthe trap into a colony, it is removed and checked for mites.

[0094] Factors such as mite population density, queen states, andenvironmental factors such as seasonality will influence the response.The amount of 2-heptanone in a particular set of circumstances that willprovide release rates within an effective attractant range can bereadily determined by a dose response test as described in Example 7,below.

[0095] Controlled release of the attractant may also be affected in partthrough the addition of an extender as known in the art, which willreduce the rate of volatilization of the attractant out of thedispenser.

[0096] Other compounds and materials may be added to a formulation,lure, bait or trap provided they do not substantially interfere with theattractancy of the attractant of the invention. Such materials includecarriers, extenders, antioxidants, ultraviolet light absorbers,pigments, dyes, fillers, blowing agents, plasticizers, other resinmodifying agents and mixtures thereof. Whether or not an additivesubstantially interferes with the attractant activity, can be determinedby standard test formats, involving direct comparisons of efficacy of2-heptanone without an added compound and 2-heptanone with an addedcompound. Reductions in attractancy, such as reduced captures of mitesin traps baited with the attractant with the additive, may be determinedwith standard statistical analyses.

[0097] The attractant of the invention may be used as a detecting agent,surveying agent, monitoring agent, or control agent for parasitic mitesof honey bees. Conveniently, the attractant is dispensed within oradjacent to a trapping means to attract and trap mites. A trappingsystem includes a trapping means and a dispenser means located withinthe trapping means which provides an effective amount of 2-heptanone. Atrapping means is any device for catching insects, particularly,parasitic mites of honey bees such as Varroa mites. These include forexample, a sticky or pitfall trap. A sticky board for use to detectinfestations of bees by organisms such as mites is described in U.S.Pat. No. 4,867,731.

[0098] The attractant of the invention is useful for control of miteswhen used in concert with other control means, such as by (a) capturingthe mites in traps as discussed above, (b) by capturing mites in a trapand killing the attracted mites, for example, by means of a drowningsolution or use of a pesticide for mites without endangering bees orcontaminating honey and wax.

[0099] Use of 2-Heptanone to Control Hive Invading Pests

[0100] The invention also relates to the use of 2-heptanone to controlhive invading pests from invading or infesting honey bee hives or otherselected areas where the presence of hive invaders is to be prevented orreduced.

[0101] One of the most damaging pests to the bee industry is the greaterwax moth (Galleria mellonella Linneaeus (Lepidoptera: Pyralidae)). Themoth enters the hive at night and deposits its eggs inside, usually increvices. The eggs hatch into larvae (caterpillar stage) and burrow intothe beeswax comb and destroy the wax combs if not discovered by thebeekeeper in time. Also, the business of beekeeping frequently requiresthe storage of empty hives and combs, particularly over winter. Usuallythe hives and combs are stored in sheds or warehouses where they arevulnerable to attack by wax moths. The wax moth is viewed by beekeepersas a major pest inflicting substantial economic losses annually,particularly in those states with mild winters. Registrations forpesticides previously used to control wax moths have been withdrawn byEPA. Hence, currently there is no registered means of controlling thispest.

[0102] Without being bound by theory, it is believed that the2-heptanone confuses the signal used by the hive invader, e.g., waxmoth, to key on a honey bee colony, and disrupts the invading behavior.Thus, application of a hive invader-controlling amount of 2-heptanone toan area prevents the target hive invader from entering the area orreduces or minimizes the numbers of hive invaders entering the area.

[0103] In this embodiment, control is carried out directly using2-heptanone. An effective hive invader-controlling amount of 2-heptanoneis provided in an area where hive invaders are to be controlled, forexample, bee hives, empty or stored bee equipment (bee hives withcombs), bee colonies, bee brood, stored honey and pollen, wax combs,hives, or wooden hive parts and the like.

[0104] In this aspect, the term control means exposing an area to ortreating an area with an effective amount of 2-heptanone, that is, anamount which is effective to prevent hive invaders from entering an areawhere they are to be controlled, or reduces or minimizes the numbers ofhive invaders entering the area. An effective hive invader-controllingamount is determined as that quantity of 2-heptanone emitted from aformulation or dispenser holding 2-heptanone that is sufficient toaccomplish the aforenamed control activities. The amount of 2-heptanonein a particular set of circumstances or for control of a particular hiveinvading pest can be determined by a dose response test. Control may becarried out using dispensers discussed in detail above. As described inthe Example below, controlled release formulations are effective tocontrol the wax moth in bee hives and wax combs. The devices can beplaced on the top board of honey bee colonies, between brood frames, orthe bottom board. Combinations of the foregoing are also contemplated bythis invention.

[0105] The lesser wax moth (Achroia grisella), though less destructivethan the greater wax moth, has similar habits and can be controlled inthe same way as described above for the greater wax moth.

[0106] It is within the compass of the invention to control other hiveinvading pests, including but not limited to, small hive beetles, ants,and Tropilaelaps, using the procedures described above.

[0107] The small hive beetle (Aethina tumida), native to Africa, wasaccidentally introduced into the United States in the early 1990's.Since then it has spread to several eastern states and continues toexpand its range. This destructive pest chews wax combs and feeds on thehoney stored therein, rapidly reducing the colony to a soggy, stickymess. There are no known natural enemies of the small hive beetle in theUnited States. One organophosphate insecticide (coumaphos) is registeredfor control.

[0108] Several species of ants (Formicidae) invade honey bee coloniesand interact with them in many ways. Some consume the wood the hive ismade of. Others consume honey, pollen or brood, and even adult bees.They weaken and destroy colonies. The need to control ants in honey beecolonies is widely recognized, however, there are few effectivetreatments.

[0109] The parasitic mite Tropilaelaps (Tropilaelaps clarae) iscurrently found throughout Southeast Asia, and parts of Africa, China,India, eastern Europe. It is anticipated that this species will beaccidentally introduced into the United States within the next 5 to 15years. These relatively large mites feed on bee brood. Damage tocolonies is usually rapid and severe following initial infestation.Effective treatment is currently limited to cumbersome managementstrategies.

EXAMPLES

[0110] The following examples are intended only to further illustratethe invention and are not intended to limit the scope of the inventionwhich is defined by the claims.

Example 1

[0111] This example describes petri dish bioassays for miticidalactivity.

[0112] 2-Heptanone was tested for miticidal activity in two separatesets of bioassays. In the petri dish bioassays we exposed free-runningadult mites to volatiles from 2-heptanone either in pure form or in anacrylic block dispenser (as described in a further experiment, below)and measured mortality over time. Mites were placed in petri dishes(5-10 mites/dish, 4 dishes per treatment and 4 dishes for controls) thatwere lined on the bottom with damp tissue paper covered with Parafilm®.Holes were punched in the Parafilm® using an insect pin to create ahumid environment in the petri dish. A piece of filter paper the samesize as the petri dish was placed on top of the Parafilm® sheet. In thefirst petri dish bioassay, 40 μl of pure 2-heptanone was placed on apiece of filter paper on the lid of the petri dish (treatment dishes).The Control dishes were set up as described above but nothing was addedto the piece of filter paper fastened to the lid of the petri dish. Thepetri dishes were placed in separate incubators for treatments andControls. Incubator temperatures were maintained at 33-34.4° C. In lessthan 2 hrs., all the mites were dead in the dishes containing the2-heptanone compared with 0% mortality in the controls. In the secondbioassay 0.1 g of a 30% concentration of 2-heptanone in acrylic blockmade as in Example 6 was placed in a plastic weighing boat in the centerof petri dishes prepared as described above. Control petri dishes wereprepared identically to those containing the acrylic block, but theweighing boat in the center of the dish was empty. In the petri dishescontaining the 2-heptanone in the acrylic block dispenser, 90% of themites were dead in 4 hrs. compared with 6% mortality in the controls.

[0113] In another series of tests, 2-heptanone, 98+% (Sigma-Aldrich);3-heptanone (Fluka Chemika); 4-heptanone (Fluka Chemika); 2-hexanone;2-octanone, 98% (Aldrich Chemical Co.); 1-heptanol (Fluka Chemika, 99.5%GC), and heptaldehyde, 95% (Aldrich Chemical Co.) were tested in petridish bioassays wherein free-running adult mites were exposed to 40 μl ofpure compound placed on filter paper as described above. The testconditions and Controls were as described above. The results are shownin Table 1, below. TABLE 1 Miticidal Activity in Petri Dish BioassaysExposure % Mortality Treatment Time (hrs) Treatment Control 2-Heptanone1 100 0 2 93 0 3-Heptanone 2 100 0 4-Heptanone 1 100 0 2 100 02-Hexanone 1 100 0 2-Octanone 1 100 0 1-Heptanol 1 65 0 2 75 10.5Heptaldehyde 1 100 0 2 100 10.5

[0114] In another series of tests, acetone (Mallinckrodt); 2-butanone,99.5+%, HPLC grade (Aldrich, Chemical Co.); ethyl butyrate, 99% (AldrichChemical Co.); benzaldehyde (Sigma Chemical Co.), and D-limonene (SigmaChemical Co. Catalog No. 12129) were tested in petri dish bioassayswherein free-running adult mites were exposed to 40 μl of pure compoundplaced on filter paper as described above. The test conditions andControls were as described above. The results are shown in Table 2,below. TABLE 2 Miticidal Activity in Petri Dish Bioassays ExposureAverage % Mortality ± S.E.* Treatment Time (hrs) Treatment ControlAcetone 1   48 ± 8.65 0 2 81.2 ± 12.0 12.5 ± 7.22 2-Butanone 1 87.5 ±7.22 0 2 77.3 ± 15.6 12.5 ± 7.22 Ethyl Butyrate 1 81.2 ± 12.0 0 2 81.2 ±12.0 0 Benzaldehyde 1   55 ± 12.6 0 2 100 0 d-Limonene 1 5.0 ± 5.0 0 255.0 ± 9.57 0

Example 2

[0115] In these experiments 2-heptanone was tested for miticidalactivity in colonies.

[0116] This set of bioassays was conducted in 5-frame colonies. Wetested miticidal activity of a 30% concentration of 2-heptanone in anacrylic block dispenser. Four colonies were established for each of thefollowing treatments: 2-heptanone, Apistan® (a commercially availablemiticide for control of Varroa), and no treatment (i.e., Controls).Before each treatment, counts were made of the number of mites thatdropped on to Dewill® Varroa Mite Detector Inserts (hereinafter denotedas “sticky boards”) placed at the bottom of colonies for one week(pre-treatment counts). A new sticky board then was inserted at thebottom of each colony, and either a single acrylic block dispenser with30% 2-heptanone made as in Example 6 (surface area=28.3 cm²) or a singleApistan® strip was placed between the frames. The Controls comprised notreatment. The number of mites on the sticky boards was counted weeklyfor 3-4 weeks afterwards (treatment interval). New sticky boards wereinserted at the beginning of each week. At the end of the test period,the 2-heptanone and Apistan® were removed, and a new sticky board wasinserted on the bottom of the colonies. A new Apistan® strip then wasplaced in the colonies to kill any remaining mites (post-treatmentinterval). The number of mites on the sticky board was counted after thecolony was exposed to the new Apistan® strip for one week. The %Reduction in the Varroa Population was estimated by summing all themites counted on the sticky board during the 3 or 4 week treatmentinterval and dividing it by the total number of mites counted for thecolony during both the treatment and post-treatment intervals. Theresults of two bioassays that followed this protocol are shown below inTable 3. TABLE 3 The percentage of Varroa mite killed by exposure to2-heptanone, fluvalinate (Apistan ®) or no treatment (Control) SampleSize % Reduction in Varroa Mite Treatment Trial (# of colonies)Population ± S.E. 2-Heptanone 1 4 61.8 ± 8.5 a Apistan ® 1 4 94.1 ± 1.5b Control 1 4 42.2 ± 7.0 c 2-Heptanone 2 3  73.6 ± 13.7 a Apistan ® 2 494.1 ± 1.2 b Control 2 4 43.6 ± 4.2 c

[0117] In Trial-1, 2-heptanone was less effective then Apistan® but moreeffective than the Control according to a Fishers Least Significantdifference test (critical value=20.5 for α=0.05). When we removed theacrylic strips from the colony after three weeks, we could still detectthe odor of 2-heptanone. We repeated the test with new colonies, and inthis trial (Trial-2) left the 2-heptanone and Apistan® strips in thecolony for 4 weeks. We then placed the post-treatment Apistan® strips inthe colonies. In Trial-2, 2-heptanone was not as effective as Apistan®but was more effective than the Control according to a Fishers LeastSignificant difference test (critical value=5.6 for α=0.05).

[0118] This test (Trial 3) was carried out in 5-frame colonies asdescribed above except for the following changes. In this test,2-heptanone was released using a controlled release strip as describedin U.S. Pat. No. 5,750,129. In brief, the controlled release strip wascomposed of a polymer sleeve 20 cm long and 5 cm wide in which apolyurethane core containing 60 g 2-heptanone was sealed. The2-heptanone strip was placed on the top of the frame (top bar). Thecontrolled diffusion of 2-heptanone through the membrane occurred over aperiod of 3 weeks. A new 2-heptanone controlled release strip was placedon the top bar after 3 weeks. The Apistan® strip was placed between theframes as described above. The Control comprised no treatment. Theresults are shown in the Table 4, below. TABLE 4 The percentage ofVarroa mite killed by exposure to 2-heptanone, fluvalinate (Apistan ®)or no treatment (Control) Sample Size % Reduction in Varroa MitePopulation Treatment (# of colonies) S.E. 2-Heptanone 4 87.05 ± 4.4 aApistan ® 4 92.40 ± 3.3 a Control 4 69.35 ± 4.8 b

Example 3

[0119] The following example describes tests to determine the influenceof 2-heptanone on honey bee colony behavior.

[0120] Tests were conducted with 2-heptanone in pure form and in a 30%concentration in acrylic block made as in Example 6 to determine itseffects on the honey bee (Apis mellifera L.) colony behavior. Atwo-frame observation hive (the hive had one frame on the bottom withadult worker bees and brood and a top frame with adult worker bees andhoney) with a laying queen was used for the study. When we placed apiece of filter paper containing 40 μl of pure 2-heptanone in theobservation colony, all the bees moved off of the frame and began to fan(i.e., move their wings up and down) vigorously. Within 4 hours the2-heptanone had evaporated, and the bees returned to the frame, and thefanning stopped.

[0121] To test the effects of 2-heptanone in acrylic blocks, we againconducted studies using two frame observation hives arranged asdescribed above. For one week prior to inserting the acrylic blocks, wemeasured queen ovipositions, and the number of workers bees leaving thehive per 5-minute interval in four observation colonies. We also countedthe number of workers in a court surrounding the queen once during the5-minute interval. We then added one acrylic block dispenser with asurface area of 8.04 cm² containing a 30% concentration of 2-heptanone.Two observation colonies contained the acrylic blocks with 2-heptanoneand two did not. The latter served as Controls.

[0122] 2-Heptanone had no immediate effect on the bees on the frameswhen we inserted it into the observation hive. The aroma of 2-heptanonecould be detected emanating from the colony, so we knew it was beingreleased in the colony. The number of worker bees forming the queen'scourt and the number of ovipositions per 5-minute interval did notdiffer in the treatment colonies after 2-heptanone was added to the hive(Table 5). Foraging activity was slightly higher after 2-heptanone wasadded as determined by a Student's t-test. TABLE 5 The average number ofworker bees in the court surrounding the queen, ovipositions by thequeen, and foragers leaving the hive per 5-minute interval in coloniesbefore and after 2-heptanone was added to the hive. Control colonies didnot have 2-heptanone added at any time during the study. PretreatmentPost-Treatment Colony Workers Foragers Workers Foragers Type in courtOvipositions leaving in court Ovipositions leaving Control 12.7 2.8 18.39.3 3.5 18.3 2-Heptanone 10.2 5.3 16.1 10.2 5.3 23.6*

Example 4

[0123] This experiment shows the Minimum Level Treatment Effectiveness.

[0124] Determination of the minimum amount of 2-heptanone effective inmite mortality in our bioassay was performed according to the followingprocedure: 3 Kimwipes® were placed on the bottom of petri dishes andslightly moistened with water using a disposable 1 ml pipette. A cutsection of Parafilm® M was placed over the moistened Kimwipes® andstretched up and over the outer edge of the petri dish bottom. TheParafilm® stretched over the Kimwipes® was then repeatedly pierced witha small dissecting insect pin to create a humid environment in the petridish. A single Whatman® Filter paper (9.0 cm) was placed on top of thepierced Parafilm® area and pushed down to create space between thefilter paper and the petri dish lid. Six to eight mites and two beelarvae were placed on top of the filter paper in each petri dish. Asmall strip of filter paper was taped to the inside top of each petridish. Four groups of two petri dishes were labeled according to the typeof treatment. The treatments were: 1 μL 2-heptanone; 5 μL 2-heptanone;10 μL 2-heptanone; and Control (no treatment). All eight dishes werethen placed in incubators set to maintain a temperature of 33-34.4° C.Our results were as follows: TABLE 6 Threshold Concentration for2-Heptanone (Low) % Mortality % Mortality % Mortality % MortalityTreatment Dish # 1 hour 17.5 hours 25.5 hours 40.5 hours Control 1 0%20% 25% 25% 2 0% 25% 25% 25% 1 μL 1 16%  33% 66% 66% 2 0%  0% 50% 50% 5μL 1 100%  100%  100%  100%  2 17%  33% 33% 68% 10 μL 1 17%  66% 66% 66%2 0% 40% 50% 60%

[0125] These results indicate that at higher concentrations, 2-heptanonehas a knockdown actions on the mites. In our 1-hour observations wethought the mites were dead but they were merely inanimate. Over time as2-heptanone evaporated the mites became active again.

[0126] In our second bioassay, the same protocol was applied; however wesought to investigate if lower amounts of 2-heptanone could be aseffective. In this trial, amounts of 0.5 and 1 μL of 2-heptanone wereused. After 17.5 hours exposure to 2-heptanone, our results were asfollows: TABLE 7 Threshold Concentration for 2-Heptanone (Low) # of DeadMites/ Treatment Dish # Total Mites % Mortality Control 1 1/5 20% 2 3/560% 3 0/6  0% 4 1/6 17% 0.5 μL 2-heptanone 1 1/6 17% 2 1/5 20% 3 2/6 34%4 0/5  0% 1 μL 2-heptanone 1 3/6 50% 2 0/5  0% 3 1/4 25% 4 1/5 20%

[0127] There was no significant difference in the average % mortalityamong the different treatments as determined by an F-test (F=0.12,df=2,9; p>0.05).

[0128] Conclusion:

[0129] Based on our results, we believe that 1 μL is the minimum amountof 2-heptanone that will cause mite mortality. This corresponds to aconcentration of 1800 ppm v/v or 0.18%.

Example 5

[0130] This experiment examines the maximum levels of 2-heptanonewithout negatively impacting bee behavior.

[0131] A test was conducted with 2-heptanone in pure form to determineits effects on honey bee (Apis mellifera L.) colony behavior andascertain if there was a maximum dose of 2-heptanone that could induce acolony to leave their hive. Two full sized frames were taken from acolony in our local apiary and placed inside a two-frame observationhive having a total volume of 8565 cm ³. These two frames containedadult worker bees only and were queenless. A 2-day period when notesting occurred was included to allow for colony adjustment to the newdwellings. The observation hive was set up inside a greenhouse with a2.5 cm tube connected from the side of the observation hive to the sidewall of the greenhouse. This tube served as an exit and entrance for theworkers.

[0132] We fastened a strip of filter paper measuring 7.6 cm by 2.5 cm tothe top inside frame of the observation colony with a thumbtack andapplied an initial 100 μL of pure 2-heptanone to the filter paper andobserved worker behavior. Subsequent applications of 40 μL of2-heptanone were applied every 4 minutes with observations of workerbehavior documented. Ambient temperature at the time of the observationswas 23° C.

[0133] After 24 minutes exposure time, we removed the initial filterpaper and replaced it with a new filter paper strip measuring 5 cm by 15cm and placed a new amount of 340 μL of 2-heptanone and resumed theapplication pattern of 40 μL ever 4 minutes until 460 μL was reached. Atthis point, we again replaced the filter paper with a new filter paperstrip measuring 5 cm by 15 cm and saturated this final piece with 460 μLof 2-heptanone.

[0134] Our experiment ended when 500 μL of pure 2-heptanone was thetotal amount on our filter paper. Concentration of 500 μL of 2-heptanonein the two-frame observation hive was calculated to be 2700 ppm v/v or0.27%.

[0135] Results.

[0136] With each 40 μL addition at the various intervals, the colony'sinitial reaction was movement away from the source of 2-heptanone(filter paper piece). However, after a period of 3-4 minutes, workersreturned and began to walk on the strip and in the immediate area of thestrip. When strips with highest concentration were placed within thecolony (100 μL, 340 μL, 460 μL), workers responded by running throughoutthe observation hive or moving towards the exit tube and exiting theobservation hive to the outside. At no time however, did all bees fillyor permanently exit the observation hive. Many of the workers filteredtowards the exit tube, but never were there more than 100-150 beesoutside the exit of the observation colony.

[0137] We believe that with each additional dosage of 2-heptanone,worker bees became acclimated to the 2-heptanone.

Example 6

[0138] This example shows how the acrylic blocks described above weremade.

[0139] Blocks having the following composition were made: isobornylmethacrylate: 50% tripropyleneglycol diacrylate  6% 2-heptanone: 44%

[0140] Polymerization was triggered with 2.9% benzoyl peroxide and 1.1%dimethylparatoluidine. Each block weighed approximately 30 grams (Diam.60 mm, Height 10 mm). The blocks were molded in an aluminum cup.

[0141] However, it was found, because of a packaging problem, that theblocks lost some active ingredient during transportation and had only30% 2-heptanone. To avoid any further loss of active ingredient, theunused samples were stored at all times in a refrigerator at 4° C. Thevalue of 30% 2-heptanone was kept for further processing of the data.The rate at which 2-heptanone was released from the blocks was measured.The experimental procedure consisted in weighing the acrylic blocks leftin their aluminum mold at regular intervals. The blocks were kept in anincubator set at 34° C. (temperature of the center of the brood nest ina honey bee hive). It is noteworthy that the incubator is not air tightand that the incubator glass door had to be opened to retrieve thesample to be weighed. An Isotemp Incubator from Fisher Scientific wasused. The incubator has a measured volume of 71 liters. The samples weremonitored for 44 days.

[0142] The release rate data at 34° C. (Table 8) shows an initial strongburst of 2-heptanone which rapidly decreases in the first 10 days tostabilize at a lower but steadier release for the following 35 days.TABLE 8 % 2-Heptanone loss Day (initial conc. 30%) Daily release inpercentage 0 0 0 1 29.13 29.13 2 38.76 9.63 3 49.04 10.28 4 55.72 6.68 768.85 4.38 8 71.82 2.97 9 74.7 2.88 10 76.09 1.39 11 77.88 1.79 14 82.311.48 15 83.43 1.12 16 84.39 0.96 18 86.24 0.92 21 88.43 0.73 24 89.980.52 30 92.36 0.4 44 95.21 0.2

Example 7

[0143] These experiments evaluate 2-heptanone as an Attractant forVarroa Mites.

[0144] We conducted bioassays to determine if 2-heptanone attractsVarroa mites. In the first bioassay, we exposed free-running adult mitesto volatiles from a 20% 2-heptanone in a gel formulation. Mites wereplaced in petri dishes (5-10 mites/dish, 4 dishes per treatment and 4dishes for controls) that were lined on the bottom with damp tissuepaper covered with Parafilm®. Holes were punched in the Parafilm® (usingan insect pin to create a humid environment in the petri dish. A pieceof filter paper the same size as the petri dish was placed on top of theParafilm® sheet. 40 μl of 20% 2-heptanone in a gel formulation was addedto a weighing boat located in the center of the petri dish. Controldishes were set up as described above with empty weighing boats in thecenter. Within 24 hours 27.3% of the mites were found in the gelformulation of 2-heptanone located in the weighing boat in the center ofthe petri dish. No mites were found in the weighing boat in the Controldish. The bioassay was repeated using the same procedure as describedabove. In the second replicate, 16.7% of the mites were found in the gelformulation of 2-heptanone located in the weighing boat in the center ofthe petri dish and again 0% were found in the weighing boats of theControl dishes.

[0145] In a second test to determine if 2-heptanone is an attractant, aVarroa trapping device was placed in a Varroa-infested 9-frame colonyfor 48 hrs. The device was constructed by placing a 2.54 cm² piece ofDewill® Varroa Mite Detector Insert coated with a 20% concentration of2-heptanone in a gel formulation and covered with a 0.63 cm (0.25 inch)wire mesh. A wire was placed through the top of the trap so that thetrap dangled between the frames in the colony. The device was placedbetween frames containing brood. After 48 hrs, several hundred Varroamites were found stuck in the gel on the trap behind the wire mesh.

[0146] A third bioassay to test for attractiveness of 2-heptanone toVarroa was conducted in petri dishes with a diameter of 13.97 cm and avolume of 76.64 cm³. The dishes were prepared as described previously inthe first bioassay of this Example using 20% gel formulation of2-heptanone. However, the petri dishes for this bioassay were modifiedby drilling a hole 1.6 cm in diameter into the center of the dish. Thishole was large enough to fit a 1.5 mL Eppendorf® tube which would serveas the dispenser for the 2-heptanone. The lid of each petri dish waslabeled in a “bulls eye” fashion with concentric circles at 1.27 cmincrements from the center where the Eppendorf® tube was located. Themaximum distance from the center was 6.35 cm. The Eppendorf® tube isconsidered the target. In the treatment dishes, 0.04 g of 2-heptanone inacrylic block formulation (30% concentration) was placed in theEppendorf® tube and covered with a small amount of sterile cotton.Control dishes were prepared in the same manner as the treatment dishesbut the Eppendorf® tubes contained only cotton. Mites (4-5 per dish)were placed in each dish in the area furthest from the center where theEppendorf® tube was located.

[0147] After the dishes were prepared a square section of mosquitonetting large enough to cover the top of the petri dish was placed overthe dish and secured with a rubber band. The mosquito netting was usedinstead of a lid on the dish. The mesh of the netting was wide enough toallow the 2-heptanone to volatilize, but prevented the mites fromescaping. The marked lids were placed on top of the dishes every 30minutes to document mite movement. After the measurements were made, thelids were immediately removed. The petri dishes were observed in 30minute increments, and the movement of the mites towards or away fromthe center Eppendorf® tube was documented by counting the number ofmites in each concentric circle.

[0148] After 1 hr significantly more mites (p<0.05) in the treatmentdishes were closer to the target (1.3 cm from the Eppendorf® tube withthe 2-heptanone) than the mites in the Control plate (Table 9). Moremites were in the 1.3 cm region throughout the bioassay, but thedifferences were not significant at the α=0.05 level. Mites were foundin the Eppendorf® tube target in both treatments and Controls, butremained in neither. TABLE 9 Percentages of Varroa mites variousdistances from a target source containing 2- heptanone (30%concentration in acrylic block delivery system) over a 4 hr. observationperiod. % of Varroa Various Distances from Target Source Exposure Time(cm) Treatment (hrs.) 6.3 5.1 3.8 2.5 1.3 Target 2-Heptanone 1 17.516.2* 25.0 7.5 27.5* 0 Control 1 18.7 50.6* 20.0 15.0 5.0* 2.52-Heptanone 2 10.0 15.0 12.5 20.0 32.5 7.5 Control 2 22.5 39.4 10.0 8.217.5 5.0 2-Heptanone 3 7.5 12.5 22.5 10.6 45.6 3.1 Control 3 10.6 32.518.7 13.1 19.4 0 2-Heptanone 4 5.5 18.8 20.0 13.3 42.2 0 Control 4 10.036.7 20.8 7.5 25.8 0

[0149] A fourth test of attractiveness was conducted in the petri dishesthat were prepared as described above. In this test 0.06 g of2-heptanone (30% concentration in acrylic block delivery system) wasplaced in the center target of the treatment plates. Control platescontained an empty Eppendorf® tube. The mites were placed 3.17 cm fromthe target. The mites could move closer or farther away from the target.Two treatment and two Control plates with 5 mites each were observed.During the 4 hr. observation period, significantly more mites movedwithin 1.27 cm of the target (38.4%) compared to the Controls (6.2%)(t=4.01, p=0.0015), and 9.1% moved into the target in the treatmentplates compared with 0.0% for the Controls.

Example 8

[0150] The following example describes use of 2-heptanone to control thehive invading pest, the wax moth (Galleria mellonella).

[0151] We conducted an experiment where we stored five frames with drawncomb in a nucleus colony box without bees present. In three nucleuscolony boxes we placed 1, 2 or 3 strips containing 60 grams of2-heptanone in a slow release formulation. In a fourth nucleus colonybox we placed no 2-heptanone. Within 2 weeks after the start of theexperiment, the equipment without 2-heptanone had wax moth adultspresent between the frames. The adults probably were laying eggs. After5 weeks, the equipment without 2-heptanone was totally infested with waxmoth. There still was no wax moth in any equipment containing2-heptanone.

[0152] It is understood that the foregoing detailed description is givenmerely by way of illustration and that modification and variations maybe made within, without departing from the spirit and scope of theinvention. All publications and patents cited herein are herebyincorporated by reference in their entirety.

What is claimed is:
 1. A method of controlling parasitic mites of honeybees, which comprises exposing parasitic mites to a miticidallyeffective amount of a compound, comprising: (a) a ketone of thestructure: CH₃—(CH2)_(x)—CO—(CH2)_(y)—CH₃  wherein y is 0 and x is 0 to5 or wherein y is 1 or 2 and x is 3 or 2, respectively; or (b) acompound selected from the group consisting of 1-heptanol, ethylbutyrate, benzaldehyde, heptaldehyde, and d-limonene, wherein saidmiticidal amount is effective to kill mites, to incapacitate mites suchas by disrupting neural or other physiological functions to preventessential mite functions or reproduction, or to render mites impairedsufficiently to be trapped, drowned, isolated, or otherwise removed froman area.
 2. The method of claim 1 wherein said parasitic mites areVarroa mites.
 3. The method of claim 1 wherein said controlling iscarried out by placing said effective amount of said miticidal compoundinside a honey bee hive so that vapors of said compound are distributedin the hive.
 4. The method of claim 1 wherein said effective amount ofsaid miticidal compound is dispensed by a dispensing means comprising adevice or formulation which provides controlled release, slow release orsustained release of said compound.
 5. The method of claim 4 whereinsaid parasitic mites are exposed to an effective amount of saidmiticidal compound for one or more brood cycles.
 6. The method of claim5 wherein said parasitic mites are exposed to an effective amount ofsaid miticidal compound for two or more brood cycles.
 7. The method ofclaim 1 wherein said miticidal compound is 2-heptanone.
 8. A method ofcontrolling parasitic mites of honey bees, which comprises placing in anarea where mites are to be attracted a dispenser means which provides anamount of 2-heptanone effective to attract parasitic mites.
 9. Themethod of claim 8 wherein said parasitic mites are Varroa mites.
 10. Themethod of claim 8 wherein said dispensing means comprises a device orformulation which provides controlled release, slow release or sustainedrelease of 2-heptanone.
 11. A method of controlling hive invading pestsof honey bees, which comprises placing in an area where pests are to becontrolled a dispenser means which provides an effective hiveinvader-controlling amount of 2-heptanone.
 12. The method of claim 11wherein said dispensing means comprises a device or formulation whichprovides controlled release, slow release or sustained release of2-heptanone.
 13. The method of claim 11 wherein said hive invading pestis selected from the group consisting of greater wax moth, lesser waxmoth, small hive beetle, ants, and Tropilaelaps.
 14. The method of claim11 wherein said hive invading pest is the greater wax moth, Galleriamellonella.
 15. A miticidal composition for controlling parasitic mitesof honey bees which comprises a dispenser means which provides amiticidally effective amount of a compound, comprising: (a) a ketone ofthe structure: CH₃—(CH2)_(x)—CO—(CH2)_(y)—CH₃  wherein y is 0 and x is 0to 5 or wherein y is 1 or 2 and x is 3 or 2, respectively; or (b) acompound selected from the group consisting of 1-heptanol, ethylbutyrate, benzaldehyde, heptaldehyde, and d-limonene, wherein saidmiticidal amount is effective to kill mites, to incapacitate mites suchas by disrupting neural or other physiological functions to preventessential mite functions or reproduction, or to render mites impairedsufficiently to be trapped, drowned, isolated, or otherwise removed froman area.
 16. The composition of claim 15 wherein said dispensing meanscomprises a device or formulation which provides controlled release,slow release or sustained release of said compound in a miticidallyeffective amount.
 17. The composition of claim 15 wherein saiddispensing means provides an effective amount of said miticidal compoundfor one or more brood cycles.
 18. The composition of claim 15 whereinsaid parasitic mites are Varroa mites.
 19. An attractant composition forattracting parasitic mites of honey bees, which comprises a dispensermeans which provides an amount of 2-heptanone effective to attractparasitic mites.
 20. The composition of claim 19 wherein said dispensingmeans comprises a device or formulation which provides controlledrelease, slow release or sustained release of 2-heptanone effective toattract parasitic mites of honey bees.
 21. The composition of claim 19wherein said parasitic mites are Varroa mites.
 22. A trapping system forcontrolling parasitic mites of honey bees, which comprises a trappingmeans and a dispenser means which provides an effective mite-attractingamount of 2-heptanone, wherein said dispenser means is located withinsufficient proximity to said trapping means so that mites that areattracted are trapped.
 23. A composition for controlling hive invadingpests of honey bees, which comprises a dispenser means which provides aneffective hive invader-controlling amount of 2-heptanone.
 24. Thecomposition of claim 23 wherein said dispensing means comprises a deviceor formulation which provides controlled release, slow release orsustained release of 2-heptanone effective to control hive invadingpests.
 25. The composition of claim 23 wherein hive invading pest isselected from the group consisting of greater wax moth, lesser wax moth,small hive beetle, ants, and Tropilaelaps.
 26. The compositions of claim23 wherein said hive invading pest is the greater wax moth, Galleriamellonella.